Esclapez J, Pire C, Camacho M, Bautista V, Martínez-Espinosa R M, Zafrilla B, Vegara A, Alcaraz L A, Bonete M J
División de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad de Alicante, Ap. 99, E-03080 Alicante, Spain.
Bioarray, S.L. Alicante, Spain.
J Biotechnol. 2015 Jan 10;193:100-7. doi: 10.1016/j.jbiotec.2014.11.018. Epub 2014 Nov 27.
The haloarchaeon Haloferax mediterranei is able to grow in the presence of different inorganic and organic nitrogen sources by means of the assimilatory pathway under aerobic conditions. In order to identify genes of potential importance in nitrogen metabolism and its regulation in the halophilic microorganism, we have analysed its global gene expression in three culture media with different nitrogen sources: (a) cells were grown stationary and exponentially in ammonium, (b) cells were grown exponentially in nitrate, and (c) cells were shifted to nitrogen starvation conditions. The main differences in the transcriptional profiles have been identified between the cultures with ammonium as nitrogen source and the cultures with nitrate or nitrogen starvation, supporting previous results which indicate the absence of ammonium as the factor responsible for the expression of genes involved in nitrate assimilation pathway. The results have also permitted the identification of transcriptional regulators and changes in metabolic pathways related to the catabolism and anabolism of amino acids or nucleotides. The microarray data was validated by real-time quantitative PCR on 4 selected genes involved in nitrogen metabolism. This work represents the first transcriptional profiles study related to nitrogen assimilation metabolism in extreme halophilic microorganisms using microarray technology.
嗜盐古菌地中海嗜盐嗜盐菌能够在有氧条件下通过同化途径在不同的无机和有机氮源存在下生长。为了鉴定在嗜盐微生物中氮代谢及其调控中潜在重要的基因,我们分析了其在三种不同氮源培养基中的全局基因表达:(a)细胞在铵中静止生长和指数生长,(b)细胞在硝酸盐中指数生长,以及(c)细胞转移到氮饥饿条件下。已确定以铵为氮源的培养物与以硝酸盐或氮饥饿的培养物之间转录谱的主要差异,支持先前的结果,即表明不存在铵作为负责硝酸盐同化途径相关基因表达的因素。结果还允许鉴定转录调节因子以及与氨基酸或核苷酸的分解代谢和合成代谢相关的代谢途径变化。通过对4个参与氮代谢的选定基因进行实时定量PCR验证了微阵列数据。这项工作代表了首次使用微阵列技术对极端嗜盐微生物中与氮同化代谢相关的转录谱研究。